COSMETIC COMPOSITION COMPRISING DENDROBIUM CANDIDUM FLOWER EXTRACT
20210059927 ยท 2021-03-04
Assignee
Inventors
- Byunghyun KIM (Daejeon, KR)
- Seyeon AHN (Daejeon, KR)
- So Young Lee (Daejeon, KR)
- Mu Hyun JIN (Daejeon, KR)
- Seung-Jin HWANG (Daejeon, KR)
Cpc classification
A23L33/105
HUMAN NECESSITIES
International classification
A23L33/105
HUMAN NECESSITIES
Abstract
Disclosed is a cosmetic composition including a Dendrobium candidum flower extract, and specifically relates to a cosmetic composition having an excellent skin improvement effect such as wrinkle improvement, skin brightening, skin trouble improvement, skin moisturizing, etc. Also disclosed is a quasi-drug composition and a food composition, which include a Dendrobium candidum flower extract. The cosmetic composition, which includes a Dendrobium candidum flower extract as an active ingredient, is excellent in wrinkle improvement, skin brightening, skin trouble improvement, or skin moisturizing effect. Specifically, the composition is excellent in collagen synthesis promoting effect, melanin production inhibitory effect, anti-inflammatory effect, and moisturizing effect, and has high utility as a cosmetic composition for wrinkle improvement, skin brightening, skin trouble improvement, or skin moisturizing. In addition, the composition, which comprises a Dendrobium candidum flower extract as an active ingredient, can be used as a quasi-drug composition and a food composition.
Claims
1. A method for improving skin condition, comprising: applying a cosmetic composition comprising a Dendrobium candidum flower extract to the skin of a subject.
2. The method of claim 1, wherein the improvement of skin condition is wrinkle improvement.
3. The method of claim 1, wherein the improvement of skin condition is skin brightening.
4. The method of claim 1, wherein the improvement of skin condition is skin trouble improvement.
5. The method of claim 1, wherein the improvement of skin condition is skin moisturizing.
6. The method of claim 1, wherein the cosmetic composition has collagen synthesis promoting effect, melanin production inhibitory effect, anti-inflammatory effect, or moisturizing effect.
7. A method for improving skin condition, comprising: administrating a food composition comprising a Dendrobium candidum flower extract to a subject.
8. The method of claim 7, wherein the improvement of skin condition is wrinkle improvement.
9. The method of claim 7, wherein the improvement of skin condition is skin brightening.
10. The method of claim 7, wherein the improvement of skin condition is skin trouble improvement.
11. The method of claim 7, wherein the improvement of skin condition is skin moisturizing.
12. The method of claim 7, wherein the food composition has collagen synthesis promoting effect, melanin production inhibitory effect, anti-inflammatory effect, or moisturizing effect.
Description
DETAILED DESCRIPTION
[0078] Hereinafter, the present invention will be described in more detail through examples. These examples are intended to illustrate the present invention more specifically, but the scope of the present invention is not limited to these examples.
Preparation Examples 1 to 5: Preparation of Whole Plant Extract of 5 Types of Dendrobii Claudis
<Preparation Example 1> Preparation of Whole Plant Extract of Dendrobium candidum
[0079] After drying the whole plant (flowers, leaves, stems, and roots) of Dendrobium candidum, 200 g of distilled water was added to 100 g of dry weight and extracted for 3 hours at 80 C. The obtained extract was filtered through a filter having a pore size of 0.2 m to prepare a Dendrobium candidum whole plant extract.
<Preparation Example 2> Preparation of Whole Plant Extract of Dendrobium nubile
[0080] Except that Dendrobium candidum was replaced with Dendrobium nobile, it was prepared in the same manner as in Preparation Example 1.
<Preparation Example 3> Preparation of Whole Plant Extract of Dendrobium loddigesii
[0081] Except that Dendrobium candidum was replaced with Dendrobium loddigesii, it was prepared in the same manner as in Preparation Example 1.
<Preparation Example 4> Preparation of Whole Plant Extract of Dendrobium fimbriatum
[0082] Except that Dendrobium candidum was replaced with Dendrobium fimbriatum, it was prepared in the same manner as in Preparation Example 1.
<Preparation Example 5> Preparation of Whole Plant Extract of Dendrobium chrysanthum
[0083] Except that Dendrobium candidum was replaced with Dendrobium chrysanthum, it was prepared in the same manner as in Preparation Example 1.
Preparation Examples 6 to 9: Preparation of Extract for Each Part of Dendrobium candidum
<Preparation Example 6> Preparation of Root Extract of Dendrobium candidum
[0084] Except that the whole plant of Dendrobium candidum was replaced with the root of Dendrobium candidum, it was prepared in the same manner as in Preparation Example 1.
<Preparation Example 7> Preparation of Stem Extract of Dendrobium candidum
[0085] Except that the whole plant of Dendrobium candidum was replaced with the stem of Dendrobium candidum, it was prepared in the same manner as in Preparation Example 1.
<Preparation Example 8> Preparation of Leaf Extract of Dendrobium candidum
[0086] Except that the whole plant of Dendrobium candidum was replaced with the leaf of Dendrobium candidum, it was prepared in the same manner as in Preparation Example 1.
<Preparation Example 9> Preparation of Flower Extract of Dendrobium candidum
[0087] Except that the whole plant of Dendrobium candidum was replaced with the flower of Dendrobium candidum, it was prepared in the same manner as in Preparation Example 1.
Experimental Example 1: Analysis of Synthesis Promotion Effect of Type 1 Collagen in Human-Derived Fibroblast
[0088] In order to confirm the wrinkle improvement effect through promoting the synthesis of type 1 collagen of each extract prepared in Preparation Examples 1 to 9, collagen synthesis rate was measured by adding the samples obtained in Preparation Examples 1 to 9 to the culture solution of human-derived fibroblasts. The synthesis of collagen was quantified using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme ImmunoAssay KIT). After adding the sample to a culture medium of fibroblasts (DMEM medium) at a final concentration of 0.1% and culturing for 48 hours, cultures were taken and the degree of collagen synthesis of type 1 collagen at each concentration was measured at 450 nm using a spectrophotometer with PICP EIA kit.
[0089] For comparison of the effects, the degree of collagen synthesis was measured in the same manner with respect to a culture medium (negative control group) of untreated fibroblasts and the sample (positive control) added with TGF- to a final concentration of 10 ng/mL.
[0090] Collagen synthesis rate (%) was calculated as the ratio of the collagen production amount relative to the negative control, and the results are shown in Table 1 below.
TABLE-US-00001 TABLE 1 Sample Absorbance (abs) Collagen synthesis rate (%) Negative control group 1.585 0 TGF- (10 ng/ml) 1.826 15.21 Preparation Example 1 1.763 11.23 (0.1%) Preparation Example 2 1.653 4.29 (0.1%) Preparation Example 3 1.692 6.75 (0.1%) Preparation Example 4 1.617 2.02 (0.1%) Preparation Example 5 1.628 2.71 (0.1%) Preparation Example 6 1.601 1.01 (0.1%) Preparation Example 7 1.723 8.71 (0.1%) Preparation Example 8 1.701 7.32 (0.1%) Preparation Example 9 1.892 19.37 (0.1%) (Number of repetitions = 4)
[0091] As shown in Table 1 above, when treated with a Dendrobium candidum extract (Preparation Example 1), it was confirmed that the collagen synthesis rate was the most excellent compared to cases of being treated with each of a Dendrobium nobile, Dendrobium loddigesii, Dendrobium fimbriatum, or Dendrobium chrysanthum extract (Preparation Examples 2 to 5).
[0092] In addition, when treated with an extract derived from the flower of Dendrobium candidum, it was confirmed that the collagen synthesis rate was the most excellent compared to being treated with the root, stem, or leaf extract of Dendrobium candidum (Preparation Examples 6 to 8), and this was more excellent than the treatment of the whole plant extract (Preparation Example 1) of Dendrobium candidum.
[0093] Additionally, the Dendrobium candidum flower extract (Preparation Example 9) showed a collagen synthesis effect that is equal to or better than that of applying TGF- (positive control group), which is generally known to induce collagen synthesis.
[0094] Accordingly, it was known that the Dendrobium candidum flower extract effectively promoted collagen synthesis compared to other types of Dendrobii Claulis and other part extracts of Dendrobium candidum, and it can be usefully applied for the use of skin wrinkle improvement using the same.
Experimental Example 2: Analysis of Inhibitory Effect of Melanin Production at Cellular Level
[0095] In order to confirm the brightening effect through the inhibition of melanin production of each extract prepared in Preparation Examples 1 to 9, the samples obtained in Preparation Examples 1 to 9 were added to a culture medium of mouse melanoma cells (B-16 mouse melanoma cell) to measure the total amount of melanin (Lotan R. et al., Cancer Res. 40:3345-3350, 1980). Prior to measuring the total amount of melanin, brightening evaluation was performed at non-toxic concentrations by first evaluating the toxicity of mouse melanoma cells.
[0096] The samples of Preparation Examples 1 to 9 were each added to a medium to a final concentration of 0.1% and B-16 melanoma cells were cultured for 3 days, by adding to a sample-untreated medium (negative control group), and to a medium of albutin to 100 ppm (positive control group), respectively.
[0097] Thereafter, the cells were trypsin-treated, detached from the culture vessel, and centrifuged, and melanin was extracted. The detached cells were added with 1 mL of sodium hydroxide solution (1N concentration), boiled for 10 minutes to dissolve the melanin, and the absorbance was measured at 405 nm using a spectrophotometer to measure the amount of melanin produced.
[0098] The amount of melanin was measured by the absorbance of the unit cell count (110.sup.6 cells), and the melanin production inhibition rate (%) was calculated as the total amount of melanin relative to the negative control group, and the results are shown in Table 2 below.
TABLE-US-00002 TABLE 2 Melanin production inhibition Sample Absorbance (abs) rate (%) Negative control group 0.652 0 Albutin (100 ppm) 0.436 33.13 Preparation Example 1 (0.1%) 0.523 19.79 Preparation Example 2 (0.1%) 0.615 5.67 Preparation Example 3 (0.1%) 0.623 4.45 Preparation Example 4 (0.1%) 0.596 8.59 Preparation Example 5 (0.1%) 0.633 2.91 Preparation Example 6 (0.1%) 0.614 5.83 Preparation Example 7 (0.1%) 0.571 12.42 Preparation Example 8 (0.1%) 0.581 10.89 Preparation Example 9 (0.1%) 0.413 36.66 (Number of repetitions = 4)
[0099] As shown in Table 2 above, when treated with a Dendrobium candidum extract (Preparation Example 1), it was confirmed that the melanin production inhibition rate was the most excellent compared to cases of being treated with each of a Dendrobium nobile, Dendrobium loddigesii, Dendrobium fimbriatum, or Dendrobium chrysanthum extract (Preparation Examples 2 to 5).
[0100] In addition, when treated with an extract derived from the flower of Dendrobium candidum (Preparation Example 9), it was confirmed that the melanin production inhibition rate was the most excellent compared to being treated with the root, stem, or leaf extract of Dendrobium candidum (Preparation Examples 6 to 8), and this was more excellent than the treatment of the whole plant extract (Preparation Example 1) of Dendrobium candidum.
[0101] Additionally, the Dendrobium candidum flower extract (Preparation Example 9) showed a melanin inhibitory effect that is equal to or better than that of albutin treatment (positive control group), which is generally known to inhibit melanin production.
[0102] Accordingly, it was known that the Dendrobium candidum flower extract effectively inhibited melanin production compared to other types of Dendrobii Claulis and other part extracts of Dendrobium candidum, and it can be usefully applied for the use of skin brightening using the same.
Experimental Example 3: Analysis of Inhibitory Effect of Nitric Oxide (NO) Production
[0103] In order to confirm the anti-inflammatory effect and skin trouble improvement effect of each extract prepared in Preparation Examples 1 to 9, NO production inhibitory effect was measured by the GRIESS method using RAW264.7 cell line (ATCC number: CRL-2278).
[0104] Specifically, RAW264.7 cells, which are macrophages of mice, were subcultured several times, and cultured for 24 hours after being placed in a 24-well plate such that 310.sup.5 cells were placed in one well. After exchanging with a cell medium containing the sample at a final concentration of 0.1%, an untreated medium was used as a negative group and for a positive control group, 20 ppm of L-NG-monomethylarginine (L-NMMA) was used, which is a NO-production inhibitor. 1 ppm of lipopolysaccharide was treated as a stimulus source and cultured for 24 hours. The supernatant was transferred to a 96-well plate by 100 L, and the GRIESS solution was added by 100 L to react at room temperature for 10 minutes, and the NO inhibitory effect was determined by measuring the absorbance at 540 nm.
[0105] The NO production inhibitory rate (%) was calculated as a relative ratio of the amount of reduced NO compared to the LPS-only treatment group, and the results are shown in Table 3 below.
TABLE-US-00003 TABLE 3 NO production Sample Absorbance (abs) inhibitory rate (%) Negative control group 0.043 LPS treated group 0.232 0 L-NMMA (20 ppm) 0.134 51.85 Preparation Example 1 (0.1%) 0.173 31.22 Preparation Example 2 (0.1%) 0.213 10.05 Preparation Example 3 (0.1%) 0.224 4.23 Preparation Example 4 (0.1%) 0.199 17.46 Preparation Example 5 (0.1%) 0.217 7.94 Preparation Example 6 (0.1%) 0.219 6.88 Preparation Example 7(0.1%) 0.181 26.98 Preparation Example 8 (0.1%) 0.195 19.58 Preparation Example 9 (0.1%) 0.131 53.44 (Number of repetitions = 4)
[0106] As shown in Table 3 above, when treated with a Dendrobium candidum extract (Preparation Example 1), it was confirmed that the NO production inhibition rate was the most excellent compared to cases of being treated with each of a Dendrobium nobile, Dendrobium loddigesii, Dendrobium fimbriatum, or Dendrobium chrysanthum extract (Preparation Examples 2 to 5).
[0107] In addition, when treated with an extract derived from the flower of Dendrobium candidum (Preparation Example 9), it was confirmed that the NO production inhibition rate was the most excellent compared to being treated with the root, stem, or leaf extract of Dendrobium candidum (Preparation Examples 6 to 8), and this was more excellent than the treatment of the whole plant extract (Preparation Example 1) of Dendrobium candidum.
[0108] Moreover, the Dendrobium candidum flower extract (Preparation Example 9) showed a NO production inhibitory effect that is equal to or better than that of L-NMMA treatment, which is a representative anti-inflammatory ingredient.
[0109] Accordingly, it was known that the Dendrobium candidum flower extract effectively inhibited NO production compared to other types of Dendrobii Claulis and other part extracts of Dendrobium candidum and thus had excellent anti-inflammatory effect, and it can be usefully applied for the use of skin trouble improvement using the same.
Experimental Example 4: Analysis of Gene Expression Effect Related to Moisturizing
[0110] In order to confirm the skin moisturizing effect of each extract prepared in Preparation Examples 1 to 9, gene expression related to moisturizing was measured using the cells cultured as below.
[0111] Human keratinocytes (HaCaT) were in a 6-hole cell culture plate 210.sup.5 each, and then cultured using Dulbecco's Modified Eagle's medium (DMEM, Gibco, Co.) including fetal bovine serum (FBS) and antibiotics for 24 hours.
[0112] After culturing the cells, it was replaced with DMEM, High Glucose, no Glutamine, no calcium (Gibco, USA), and then the samples obtained in Preparation Examples 1 to 9 were treated at a concentration of 0.1% for 24 hours. After the cells were recovered and washed with phosphate buffer solution (PBS), total RNA was extracted using an RNA extraction kit (RNeasy mini kit, Qiagen, Germany).
[0113] 2.5 g of RNA was quantified and reverse transcription was performed using cDNA synthesis kit (PhileKorea, Korea). Reverse transcription was performed using a Veriti 96 well Thermal Cycler (Applied Biosystems, USA), and the synthesized cDNA was used by 100 ng per reaction, and TaqMan Universal Master Mix II was used. Quantitative real time PCR (qRT-). PCR) was performed with AQP3, FLG, HAS2, and HAS3 primers (TaqMan Gene Expression Assays, Thermo Fisher, USA) that were run on a StepOnePlus Real-time PCR System (Applied Biosystems, USA).
[0114] The experimental results obtained by qRT-PCT were shown by calculating with the CT method (Livak K J, Schmittgen T D: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method, Methods, 25, 402-408(2001)) based on housekeeping gene and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). To compare effects, moisturizing gene expression was measured in the same manner for a negative control group without treating samples and a positive control group treated with EGF 10 ng/ml and Ca.sup.2+3 mM, respectively.
[0115] The results of quantifying the mRNA expression level of the treated experimental group based on the mRNA expression level of the negative control group, which was not treated, as 1.00 were shown in Table 4.
TABLE-US-00004 TABLE 4 Hyaluronic acid Hyaluronic acid Aquaporin 3 synthase 2 synthase 3 Sample (fold) (fold) (fold) Negative control 1.00 1.00 1.00 group EGF (10 ng/ml) 3.27 Ca.sup.2+ (3 mM) 2.01 3.60 Preparation Example 1.51 1.73 1.82 1 (0.1%) Preparation Example 112 1.23 1.18 2 (0.1%) Preparation Example 1.09 1.11 1.04 3 (0.1%) Preparation Example 1.15 1.13 1.09 4 (0.1%) Preparation Example 1.21 1.28 1.26 5 (0.1%) Preparation Example 1.05 1.09 1.10 6 (0.1%) Preparation Example 1.51 1.64 1.69 7 (0.1%) Preparation Example 1.42 1.53 1.56 8 (0.1%) Preparation Example 2.75 2.93 2.96 9 (0.1%) (Number of repetitions = 4)
[0116] As shown in Table 4 above, when treated with a Dendrobium candidum extract (Preparation Example 1), it was confirmed that the expression of all of genes related to skin moisturizing used in the experiment was the most excellent compared to cases of being treated with each of a Dendrobium nobile, Dendrobium loddigesii, Dendrobium fimbriatum, or Dendrobium chrysanthum extract (Preparation Examples 2 to 5).
[0117] In addition, when treated with an extract derived from the flower of Dendrobium candidum (Preparation Example 9), it was confirmed that the expression of the genes related to skin moisturizing was the most excellent compared to being treated with the root, stem, or leaf extract of Dendrobium candidum (Preparation Examples 6 to 8), and this was more excellent than the treatment of the whole plant extract (Preparation Example 1) of Dendrobium candidum.
[0118] Accordingly, it was known that the Dendrobium candidum flower extract showed excellent skin moisturizing effect compared to other types of Dendrobii Claulis and other part extracts of Dendrobium candidum, and it can be usefully applied for the use of skin moisturizing using the same.
[0119] Through the results of Experimental Examples 1 to 4 above, the Dendrobium candidum extract according to the present invention has more excellent skin improvement effect (wrinkle improvement, skin brightening, skin trouble improvement, and skin moisturizing) compared to Dendrobium nobile, Dendrobium loddigesii, Dendrobium fimbriatum, and Dendrobium chrysanthum extracts, and it was confirmed that the effect was the most excellent when using an extract obtained from the flower part in Dendrobium candidum.
[0120] Therefore, the Dendrobium candidum flower extract can be effectively used in compositions for wrinkle improvement, skin brightening, skin trouble improvement, or skin moisturizing.
[0121] From the above description, those skilled in the art will appreciate that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, the embodiments described above are to be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present invention.